Supplementary Table 1. Characteristic of patients with AMD and individuals without visual disturbances (controls) enrolled in the study.
Characteristics / Controls / AMD / pNumber / Frequency / Number / Frequency
Sex / 0.0296
Females / 82 / 0.78 / 182 / 0.65
Males / 23 / 0.22 / 91 / 0.33
Unknown / 0 / - / 6 / 0.02
Age / < 0.001
≤ 60 / 31 / 0.30 / 31 / 0.11
> 60 / 74 / 0.70 / 242 / 0.89
Mean ± SD / 68.29 ± 10.95 / 72.52 ± 8.43
Range / 50-88 / 52-93
Unknown / 0 / 6
Smoking / 0.9539
Ever / 32 / 0.41 / 68 / 0.40
Never / 46 / 0.59 / 100 / 0.60
Living environment / 0.2459
Rural / 32 / 0.38 / 50 / 0.30
Urban / 51 / 0.62 / 115 / 0.70
AMD in family / 0.001
Yes / 3 / 0.04 / 33 / 0.20
No / 81 / 0.96 / 131 / 0.80
p values for a two-sides χ2 –test, p values< 0.05 are in bold.
Supplementary Table 2. Risk of AMD associated with age, sex, tobacco smoking, living environment and family history of AMD.
Chracteristics / Controls / AMD / Crude OR (95% CI) / pNumber (frequency) / Number (frequency)
Age
from point / 1.05 (1.02-1.07) / < 0.001
from range / 7.77 (2.73-22.12) / < 0.001
Sex
Females / 82 (0.78) / 182 (0.67) / 0.56 (0.33-0.95) / 0.031
Males / 23 (0.22) / 91 (0.33) / 1.78 (1.05-3.02) / 0.031
Tobacco smoking
Yes (current, former) / 32 (0.41) / 68 (0.40) / 0.97 (0.57-1.69) / 0.935
Never / 46 (0.59) / 100 (0.60) / 1.13 (0.65-1.96) / 0.661
Living environment
Rural / 32 (0.38) / 50 (0.30) / 0.71 (0.41-1.23) / 0.217
Urban / 51 (0.62) / 115 (0.70) / 1.44 (0.83-2.51) / 0.193
AMD in family
Yes / 3 (0.04) / 33 (0.20) / 6.80 (2.02-22.90) / 0.002
No / 81 (0.96) / 131 (0.80) / 0.15 (0.05-0.52) / 0.002
OR – odds ratio; 95% CI – 95% confidence interval; p values< 0.05 along with corresponding ORs are in bold.
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Supplementary Table 3. Distribution of genotypes, frequency of alleles of the 19G>C-HMOX1 and -42+1444A>G-HMOX2 polymorphisms and odds ratio (OR) with 95% confidence interval (95% CI) in patients with AMD, the disease in its dry and wet forms and individuals without visual disturbances (controls).
Genotype/allele / Controls / AMD / Crude OR(95% CI) / Adjusted ORa
(95% CI) / Dry AMD / Crude OR
(95% CI) / Adjusted ORa
(95% CI) / Wet AMD
(95% CI) / Crude OR
(95% CI) / Adjusted ORa
(95% CI)
Number (frequency) / Number (frequency) / Number (frequency) / Number (frequency)
c.19G>C,
rs2071747 / n = 105 / n = 277 / n = 100 / n = 177
G/G / 96 (0.91) / 248 (0.90) / 0.83 (0.38-1.82) / 0.75 (0.30-1.90) / 97 (0.97) / 3.03 (0.80-11.54) / 2.60 (0.60-11.35) / 151 (0.85) / 0.57 (0.25-1.27) / 0.47 (0.18-1.24)
G/C / 8 (0.08) / 26 (0.09) / 1.26 (0.55-2.87) / 1.58 (0.60-4.13) / 3 (0.03 / 0.38 (0.10-1.46) / 0.47 (0.11-2.13) / 23 (0.13) / 1.81 (0.78-4.21) / 2.55 (0.93-7.05)
C/C / 1 (0.01) / 3 (0.01) / 1.14 (0.12-11.07) / 0.45 (0.03-7.86) / 0 (-) / - / - / 3 (0.02) / 1.79 (0.18-17.46) / 0.73 (0.04-13.02)
χ2 = 0.310; p = 0.856 / χ2 = 3.158; p = 0.206 / χ2 = 0.2.270; p =0.321
G / 200 (0.95) / 522 (0.94) / 0.84 (0.42-1.66) / 0.80 (0.35-1.83) / 197 (0.99) / 2.97 (0.83-10.67) / 2.63 (0.67-10.25) / 325 (0.92) / 0.60 (0.30-1.21) / 0.73 (0.04-13.02)
C / 10 (0.05) / 32 (0.06) / 1.20 (0.60-2.37) / 1.25 (0.55-2.85) / 3 (0.01) / 0.34 (0.09-1.21) / 0.38 (0.10-1.49) / 29 (0.08) / 1.67 (0.83-3.38) / 1.90 (0.79-4.48)
c.-42+1444A>G,
rs2270363 / n = 105 / n = 274 / n = 99 / n = 175
G/G / 58 (0.55) / 131 (0.48) / 0.74 (0.42-1.17) / 0.55 (0.30-1.04) / 33 (0.33) / 0.41 (0.23-0.72) / 0.25 (0.11-0.56) / 98 (0.56) / 1.03 (0.63-1.68) / 0.91 (0.46-1.81)
G/A / 40 (0.38) / 126 (0.46) / 1.38 (0.87-2.19) / 1.75 (0.93-3.28) / 62 (0.63) / 2.73 (1.55-4.80) / 3.74 (1.70-8.27) / 64 (0.37) / 0.94 (0.57-1.54) / 1.03 (0.50-2.15)
A/A / 7 (0.07) / 17 (0.06) / 0.93 (0.37-2.30) / 1.29 (0.38-4.25) / 4 (0.04) / 0.59 (0.17-2.08) / 1.22 (0.25-5.98) / 13 (0.07) / 1.12 (0.43-2.91) / 1.24 (0.34-4.60)
χ2 = 1.944; p = 0.378 / χ2 = 12.266; p = 0.002 / χ2 = 1.101; p = 0.095
G / 156 (0.74) / 388 (0.71) / 0.83 (0.57-1.20) / 0.65 (0.40-1.07) / 128 (0.65) / 0.57 (0.36-0.93) / 0.38 (0.20-0.75) / 260 (0.74) / 1.00 (0.68 (1.47) / 0.91 (0.54-1.55)
A / 54 (0.26) / 160 (0.29) / 1.23 (0.43-1.79) / 1.54 (0.94-2.53) / 70 (0.35) / 1.77 (1.09-2.86) / 2.59 (1.34-5.01) / 90 (0.26) / 1.01 (0.69-1.49) / 1.10 (0.65-1.87)
p values for a two-sides χ2 –test, significant p ORs are in bold.
a OR adjusted for age, sex, AMD in family, living environment and tobacco smoking.
“-“ not estimated.
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Supplementary Table 4. Distribution of genotypes, frequency of alleles of the 19G>C-HMOX1 and -42+1444A>G-HMOX2 polymorphisms and odds ratio (OR) with 95% confidence interval (95% CI) in patients with wet and dryforms of AMD.
Polymorphism / Dry AMD / Wet AMD / Crude OR(95% CI) / p / Adjusted ORa
(95% CI) / p
Genotype/allele / Number (frequency) / Number (frequency)
19G>C
G/G / 97 (0.97) / 151 (0.85) / 0.19 (0.05-0.64) / 0.007 / 0.24 (0.07-0.89) / 0.033
G/C / 3 (0.03) / 23 (0.13) / 4.83 (1.41-16.52) / 0.012 / 4.07 (1.11-14.91) / 0.034
C/C / 0 (-) / 3 (0.02) / 4.03 (0.21-78.85) / 0.358 / -
G / 197 (0.99) / 325 (0.92) / 0.19 (0.06-0.63) / 0.007 / 0.23 (0.07-0.84) / 0.025
C / 3 (0.01) / 29 (0.08) / 5.20 (1.58-17.14) / 0.007 / 4.28 (1.20-15.30) / 0.025
-42+1444A>G
G/G / 33 (0.33) / 98 (0.56) / 2.55 (1.52-4.25) / < 0.001 / 3.63 (1.76-7.50) / < 0.001
G/A / 62 (0.63) / 64 (0.37) / 0.34 (0.21-0.75) / < 0.001 / 0.25 (0.12-0.51) / < 0.001
A/A / 4 (0.04) / 13 (0.07) / 1.91 (0.60-6.01) / 0.271 / 1.61 (0.39-6.63) / 0.509
G / 128 (0.65) / 260 (0.74) / 1.69 (1.12-2.55) / 0.012 / 2.10 (1.19-3.74) / 0.010
A / 70 (0.35) / 90 (0.26) / 0.59 (0.32-0.89) / 0.013 / 0.48 (0.27-0.85) / 0.012
p values < 0.05 along with corresponding ORs are in bold.
aOR adjusted for age, sex, AMD in family, living environment and tobacco smoking.
“-“ not estimated
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Supplementary Table 5.Distribution of combined genotypes of the 19G>C-HMOX1 and -42+1444A>G-HMOX2 polymorphisms and odds ratio (OR) with 95% confidence interval (95% CI) in patients with AMD as well as its dry and wet forms and individuals without visual disturbances (controls).
Genotype / Controls (n=105) / AMD(n=272) / Crude OR
(95% CI) / Dry AMD
(n=98) / Crude OR
(95% CI) / Wet AMD
(n=174) / Crude OR
(95% CI)
Number
(frequency) / Number (frequency) / Number (frequency) / Number (frequency)
G/G-G/G / 54 (0.51) / 114 (0.42) / 0.68 (0.43-1.07) / 32 (0.33) / 0.46 (0.26-0.81) / 82 0.49 / 0.84 (0.52-1.37)
G/G-A/G / 37 (0.35) / 113 (0.41) / 1.31 (0.82-2.08) / 60 (0.61) / 2.90 (1.64-5.13) / 53 0.31 / 0.80 (0.48-1.35)
G/G-A/A / 5 (0.05) / 17 (0.06) / 1.33 (0.48-3.71) / 4 (0.04) / 0.85 (0.22-3.27) / 13 0.07 / 1.62 (0.56-4.67)
G/C-G/G / 4 (0.04) / 14 (0.05) / 1.37 (0.44-4.26) / 1 (0.01) / 0.26 (0.03-2.37) / 13 0.06 / 2.04 (0.65-6.43)
G/C-A/G / 2 (0.02) / 11 (0.04) / 2.17 (0.47-9.96) / 1 (0.01) / 0.53 (0.05-5.95) / 10 0.05 / 3.14 (0.68-14.62)
G/C-A/A / 2 (0.02) / 0 (-) / 0.08 (0.00-1.59) / 0 (-) / 0.21 (0.01-4.43) / 0 (-) / 0.12 (0.01-2.49)
C/C-G/G / 0 (-) / 2 (0.02) / 1.95 (0.09-40.96) / 0 (-) / 1.07 (0.02-54.50) / 2 0.02 / 3.06 (0.15-64.32)
C/C-A/G / 1 (0.01) / 1 (0.01) / 0.38 (0.02-6.19) / 0 (-) / 0.35 (0.01-8.78) / 1 0.01 / 0.61 (0.04-9.71)
C/C-A/A / 0 (-) / 0 (-) / 0.39 (0.01-19.64) / 0 (-) / 1.07 (0.02-54.50) / 0 (-) / 0.60 (0.01-30.69)
Significant ORs are in bold; “-“ not estimated.
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Supplementary Table 6. Distribution of combined genotypes of the 19 G>C-HMOX1 and -42+1444 A>G-HMOX2polymorphismsand odds ratio (OR) with 95% confidence interval (95% CI) in patients with wet and dryforms of AMD.
Genotype / Dry AMDn = 98 / Wet AMD
n = 174 / Crude OR
(95% CI)
Number / Frequency / Number / Frequency
G/G-G/G / 32 / 0.33 / 82 / 0.47 / 1.84 (1.10-3.08)
G/G-A/G / 60 / 0.61 / 53 / 0.30 / 0.29 (0.18-0.49)
G/G-A/A / 4 / 0.04 / 13 / 0.07 / 1.50 (0.52-4.35)
G/C-G/G / 1 / 0.01 / 13 / 0.07 / 5.91 (0.75-46.92)
G/C-A/G / 1 / 0.01 / 10 / 0.06 / 5.29 (0.66-42.40)
G/C-A/A / 0 / 0 / 0 / 0 / -
C/C-G/G / 0 / 0 / 2 / 0.01 / 2.86 (0.14-60.07)
C/C-A/G / 0 / 0 / 1 / 0.01 / 1.70 (0.07-42.21)
C/C-A/A / 0 / 0 / 0 / 0 / -
Significant ORs are bolded.
“-“ not estimated.
Supplementary Table 7.Distribution of genotypes of the -42+1444A>G-HMOX2 polymorphism stratified by age, sex and AMD family history in patients with dry AMD as compared with individuals without visual disturbances (controls).
Genotype/ allele / Controls / Dry AMD / Adjusted ORa / pNumber (frequency) / Number (frequency)
Age > 61 / n = 74 / n = 84
G/G / 43 (0.58) / 29 (0.35) / 0.25 (0.10-0.61) / 0.002
G/A / 28 (0.38) / 53 (0.63) / 4.38 (1.76-10.90) / 0.001
A/A / 3 (0.04) / 2 (0.02) / 0.57 (0.04-7.44) / 0.669
G / 114 (0.77) / 111 (0.66) / 0.34 (0.15-0.78) / 0.011
A / 34 (0.23) / 57 (0.34) / 2.88 (1.28-6.48) / 0.011
Women / n = 82 / n = 65
G/G / 46 (0.56) / 20 (0.31) / 0.20 (0.07-0.54) / 0.002
G/A / 32 (0.39) / 41 (0.63) / 3.31 (1.31-8.35) / 0.011
A/A / 4 (0.05) / 4 (0.06) / 4.71 (0.68-32.64) / 0.117
G / 124 (0.76) / 81 (0.62) / 0.26 (0.11-0.59) / 0.001
A / 40 (0.24) / 49 (0.38) / 3.92 (1.70-9.02) / 0.001
Negative family history for AMD / n = 81 / n = 52
G/G / 50 (0.62) / 18 (0.35) / 0.25 (0.11-0.57) / 0.001
G/A / 26 (0.32) / 30 (0.58) / 3.83 (1.698.68) / 0.001
A/A / 5 (0.06) / 4 (0.07) / 1.18 (0.24-5.74) / 0.836
G / 126 (0.78) / 66 (0.63) / 0.39 (0.20-0.76) / 0.006
A / 36 (0.22) / 38 (0.37) / 2.59 (1.32-5.07) / 0.006
p values < 0.05 along with corresponding ORs are in bold.
a OR adjusted for age, sex, AMD in family, living environment and tobacco smoking.
Supplementary Fig. 1. Genotypes of the 19 G>C-HMOX1 polymorphism (rs2071747) determined by the allele specific oligonucleotide PCR (ASO-PCR) and analyzed by a 3% agarose gel electrophoresis stained with ethidium bromide and viewed under UV light. Lane M displays GeneRuler ™ 100 bp molecular weight marker; lanes designated G or C show the results of amplification with primer specific to either the G or C allele, respectively. X lane shows negative control. Genotypes are indicated in the lower part of the picture.
Supplementary Fig. 2. Genotypes of the -42+1444 A>G-HMOX2 polymorphism (rs2270363) determined by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) detection and analyzed by a 8% polyacrylamide gel electrophoresis stained with ethidium bromide and viewed under UV light. Lane M shows GeneRuler ™ 100 bp molecular weight marker, lane X shows a negative control, all remaining lanes present genotypes indicated in the upper part of the picture.
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